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J. Biol. Chem., Vol. 281, Issue 33, 23297-23301, August 18, 2006

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Minireview

de FACTo Nucleosome Dynamics^*

Danny Reinberg¹ and Robert J. Sims, III²

From the Howard Hughes Medical Institute and Division of Nucleic Acids Enzymology, Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854

The factors required for the delivery of RNA polymerase II to class II promoters using naked DNA were all identified by 1998, yet their exact mechanisms of action were not fully understood in all cases, and in some instances, their precise function still remains unknown. Nonetheless, a complete understanding of the complexity of the RNA polymerase II transcription cycle necessitated the development of assays that include chromatinized DNA templates. At this time, the field was actively searching for factors that allow transcription initiation on chromatinized templates. We began studies using chromatin templates in an attempt to identify factor(s) that permit RNA polymerase II to traverse nucleosomes, i.e. that allow elongation on chromatinized DNA templates. The challenge herein was to develop an assay that directly measured the ability of transcriptionally engaged RNA polymerase II to traverse nucleosomes. This approach resulted in the isolation of FACT, a heterodimer in humans comprised of Spt16 and SSRP1. Defined functional biochemical assays corroborated genetic studies in yeast that allowed the elucidation of FACT function in vivo. Collectively, these approaches demonstrate that FACT is a factor that allows RNA polymerase II to traverse nucleosomes in vitro and in vivo by removing one H2A/H2B dimer. More recent studies using a fully defined chromatin reconstitution/transcription assay revealed that FACT activity is greatly stimulated by post-translational modification of the histone polypeptides, specifically by monoubiquitination of lysine 120 of human histone H2B.

^* This minireview will be reprinted in the 2006 Minireview Compendium, which will be available in January, 2007. The studies on FACT were supported by National Institutes of Health Grant GM37120 and by the Howard Hughes Medical Institute.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. S1 and supplemental Ref. 1.

² Supported by National Institutes of Health Grant GM71166.

¹ To whom correspondence should be addressed. Tel.: 732-235-4195; Fax: 732-235-5294; E-mail: reinbedf{at}umdnj.edu.

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